甘蓝型油菜BnaSLY1基因进化分析及功能研究  

Phylogenetic and functional analysis of the BnaSLY1 genes in Brassica napus L.

作　　者：李嘉欣 黄莹莹 吴潞梅 赵伦 易斌[1] 马朝芝[1] 涂金星[1] 沈金雄[1] 傅廷栋[1] 文静[1] LI Jia-Xin;HUANG Ying-Ying;WU Lu-Mei;ZHAO Lun;YI Bin;MA Chao-Zhi;TU Jin-Xing;SHEN Jin-Xiong;FU Ting-Dong;WEN Jing(National Key Laboratory of Crop Genetic Improvement,Huazhong Agricultural University/National Engineering Research Center of Rapeseed/Hongshan Laboratory,Wuhan 430070,Hubei,China)

机构地区：[1]华中农业大学作物遗传改良全国重点实验室/国家油菜工程技术研究中心/洪山实验室,湖北武汉430070

出　　处：《作物学报》2025年第1期44-57,共14页Acta Agronomica Sinica

基　　金：国家自然科学基金项目(31771831,31000721);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-12)资助。

摘　　要：赤霉素调控植物表皮细胞增长、茎叶伸长以及株型建成。拟南芥SLY1属于F-box蛋白,它通过靶向泛素化赤霉素信号转导路径的负向调控因子——DELLA蛋白来影响植物生长发育,然而油菜中BnaSLY1的基因功能尚未揭示。本研究对BnaSLY1进行了表达特征和进化树分析,利用CRISPR/Cas9技术创制了BnaSLY1不同拷贝数的突变体,结合RNA-Seq技术对BnaSLY1的生物学功能及其对油菜生长发育的影响进行了研究。结果表明,甘蓝型油菜Westar中有2个SLY1同源拷贝BnaA01.SLY1和BnaA06.SLY1,它们表达模式基本相同,为组成型表达基因,其蛋白定位在细胞核,且在不同的油菜品种及十字花科植物间序列保守。与对照相比,单突bnaa01sly1和bnaa06sly1开花时间推迟,株高显著降低,而双突bnasly1还表现出深绿色光叶表型,叶片厚度增加,开花期比单突进一步推迟,株高也进一步降低。RNA-Seq结果显示,双突与Westar之间的差异表达基因显著富集在生长素信号转导路径以及蜡质合成通路,多个开花时间相关基因表达也发生显著变化。本研究表明, BnaSLY1除影响植物株高和开花时间等生长发育进程,还影响表皮蜡质合成,为探索赤霉素信号转导路径在甘蓝型油菜生长发育中的重要作用奠定了理论基础。Gibberellins regulate plant epidermal cell growth,stem and leaf expansion,and plant architecture.In Arabidopsis,SLY1 encodes an F-box protein that modulates plant growth by targeting the negative regulator of GA signaling,the DELLA pro-tein,for ubiquitination and subsequent degradation.However,the function of BnaSLY1 in Brassica napus has not been previously revealed.In this study,we characterized the expression patterns and performed a phylogenetic analysis of BnaSLY1.Using CRISPR/Cas9 technology,we generated mutants with different copy numbers of BnaSLY1.By integrating RNA-Seq analysis,we investigated the biological functions of BnaSLY1 and its impact on the growth and development of Brassica napus.Our results showed that there are two copies of SLY1 in Brassica napus,with similar expression patterns and constitutive expression.The protein is localized in the nucleus and is highly conserved among different varieties of rapeseed and cruciferous plants.Pheno-typic analysis of mutants revealed that,compared to the control,single mutants bnaa01sly1 and bnaa06sly1 exhibited delayed flowering and significantly reduced plant height,while the double mutant bnasly1 showed a dark green leaf phenotype,increased leaf thickness,further delayed flowering,and further reduced plant height.RNA-Seq analysis between Westar and bnasly1 showed significant enrichment of differentially expressed genes in the auxin signaling pathway and wax biosynthesis pathway,with several flowering time-related genes showing significant expression changes.This study demonstrates that BnaSLY1 not only influences plant height and flowering time but also affects epidermal wax synthesis,thereby laying a theoretical foundation for exploring the crucial role of the GA signaling pathway in the growth and development of Brasscia napus.

关 键 词：甘蓝型油菜 BnaSLY1 赤霉素 株型 蜡质合成 开花时间 RNA-SEQ 

分 类 号：S565.4[农业科学—作物学]